A sensitive and versatile in situ hybridization protocol for gene expression analysis in developing amniote brains

Pei Shan Hou, Takuma Kumamoto, Carina Hanashima*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

4 Scopus citations

Abstract

The detection of specific RNA molecules in embryonic tissues has wide research applications including studying gene expression dynamics in brain development and evolution. Recent advances in sequencing technologies have introduced new animal models to explore the molecular principles underlying the assembly and diversification of brain circuits between different amniote species. Here, we provide a step-by-step protocol for a versatile in situ hybridization method that is immediately applicable to a range of amniote embryos including zebra finch and Madagascar ground gecko, two new model organisms that have rapidly emerged for comparative brain studies over recent years. The sensitive detection of transcripts from low to high abundance expression range using the same platform enables direct comparison of gene of interest among different amniotes, providing high-resolution spatiotemporal information of gene expression to dissect the molecular principles underlying brain evolution.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages319-334
Number of pages16
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1650
ISSN (Print)1064-3745

Keywords

  • Amniote embryos
  • Brain development
  • Dual color in situ hybridization
  • Evolution
  • Gecko
  • Pallium
  • Zebra finch

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